Team: Biology and biotechnology of grapevine viruses
Team leader: Christophe RITZENTHALER
PhD supervisor: Christophe RITZENTHALER
Email: ritzenth@unistra.fr
RNA viruses establish membrane-bound viral factories in host cells to replicate their genomes, relying on a complex interplay of viral and host-encoded proteins. This process generates double-stranded RNA (dsRNA) intermediates, key elements in antiviral defense. By focusing on dsRNA in healthy and virus-infected Arabidopsis, we have identified a small set of host proteins that may function as either pro- or antiviral factors. Among these, one highly conserved protein stands out. Despite its evolutionary conservation in plants, its function remains unknown.
Our recent biochemical and structural studies, employing site-directed mutagenesis, X-ray crystallography and Cryo-EM, reveal that this protein has exceptional dsRNA-binding properties, potentially linked to plant innate immunity. This PhD project seeks to unravel the biological role of this protein under normal conditions and during viral infections. Using CRISPR-Cas9-generated mutants, phenotypic screens will be conducted under various environmental conditions, alongside testing viruses with or without silencing suppression activity. RNA-seq and small RNA profiling will be used to investigate changes in gene expression and small RNA pathways.
To explore molecular mechanisms, the project will integrate advanced techniques, super resolution microscopy, co-immunoprecipitation, and purification of native dsRNA-protein complexes for structural studies. This interdisciplinary project offers a unique opportunity to delve into plant immunity and antiviral defense using state-of-the-art molecular, genetic, and structural biology approaches.
Keywords: RNA virus replication, Host-virus interactions, dsRNA-binding proteins, plant virology
Relevant publications:
– Incarbone, M., Clavel, M., Monsion, B., Kuhn, L., Scheer, H., Vantard, E., Poignavent, V., Dunoyer, P., Genschik, P., and Ritzenthaler, C. (2021). Immunocapture of dsRNA-bound proteins provides insight into Tobacco rattle virus replication complexes and reveals Arabidopsis DRB2 to be a wide-spectrum antiviral effector. Plant Cell 33, 3402-3420. https://www.ncbi.nlm.nih.gov/pubmed/34436604
– Incarbone, M., Scheer, H., Hily, J.M., Kuhn, L., Erhardt, M., Dunoyer, P., Altenbach, D., and Ritzenthaler, C. (2020). Characterization of a DCL2-Insensitive Tomato Bushy Stunt Virus Isolate Infecting Arabidopsis thaliana. Viruses 12, 1121. https://www.ncbi.nlm.nih.gov/pubmed/33023227
– Monsion, B., Incarbone, M., Hleibieh, K., Poignavent, V., Ghannam, A., Dunoyer, P., Daeffler, L., Tilsner, J., and Ritzenthaler, C. (2018). Efficient Detection of Long dsRNA in Vitro and in Vivo Using the dsRNA Binding Domain from FHV B2 Protein. Front Plant Sci 9, 70. https://www.ncbi.nlm.nih.gov/pubmed/29449856
Financial support of this subject is guaranteed by the Research Cluster / LabEx NetRNA
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📅 Application Deadline: January 2025
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